Fibroblast-like cells change gene expression of bone remodelling markers in transwell cultures
Abstract Introduction Periprosthetic fibroblast-like cells (PPFs) play an important role in aseptic loosening of arthroplasties. Various studies have examined PPF behavior in monolayer culture systems. However, the periprosthetic tissue is a three-dimensional (3D) mesh, which allows the cells to int...
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| Format: | Article |
| Language: | English |
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BMC
2020-10-01
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| Series: | European Journal of Medical Research |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s40001-020-00453-y |
| _version_ | 1819115493637750784 |
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| author | Eliza S. Hartmann Sabine Schluessel Miriam I. Köhler Felicitas Beck Julia I. Redeker Burkhard Summer Veronika Schönitzer Andreas Fottner Susanne Mayer-Wagner |
| author_facet | Eliza S. Hartmann Sabine Schluessel Miriam I. Köhler Felicitas Beck Julia I. Redeker Burkhard Summer Veronika Schönitzer Andreas Fottner Susanne Mayer-Wagner |
| author_sort | Eliza S. Hartmann |
| collection | DOAJ |
| description | Abstract Introduction Periprosthetic fibroblast-like cells (PPFs) play an important role in aseptic loosening of arthroplasties. Various studies have examined PPF behavior in monolayer culture systems. However, the periprosthetic tissue is a three-dimensional (3D) mesh, which allows the cells to interact in a multidirectional way. The expression of bone remodeling markers of fibroblast-like cells in a multilayer environment changes significantly versus monolayer cultures without the addition of particles or cytokine stimulation. Gene expression of bone remodeling markers was therefore compared in fibroblast-like cells from different origins and dermal fibroblasts under transwell culture conditions versus monolayer cultures. Methods PPFs from periprosthetic tissues (n = 12), osteoarthritic (OA) synovial fibroblast-like cells (SFs) (n = 6), and dermal fibroblasts (DFs) were cultured in monolayer (density 5.5 × 103/cm2) or multilayer cultures (density 8.5 × 105/cm2) for 10 or 21 days. Cultures were examined via histology, TRAP staining, immunohistochemistry (anti-S100a4), and quantitative real-time PCR. Results Fibroblast-like cells (PPFs/SFs) and dermal fibroblasts significantly increased the expression of RANKL and significantly decreased the expression of ALP, COL1A1, and OPG in multilayer cultures. PPFs and SFs in multilayer cultures further showed a higher expression of cathepsin K, MMP-13, and TNF-α. In multilayer PPF cultures, the mRNA level of TRAP was also found to be significantly increased. Conclusion The multilayer cultures are able to induce significant expression changes in fibroblast-like cells depending on the nature of cellular origin without the addition of any further stimulus. This system might be a useful tool to get more in vivo like results regarding fibroblast-like cell cultures. |
| first_indexed | 2024-12-22T05:02:04Z |
| format | Article |
| id | doaj.art-53bc3cf378a64d019c93ee2ac412830b |
| institution | Directory Open Access Journal |
| issn | 2047-783X |
| language | English |
| last_indexed | 2024-12-22T05:02:04Z |
| publishDate | 2020-10-01 |
| publisher | BMC |
| record_format | Article |
| series | European Journal of Medical Research |
| spelling | doaj.art-53bc3cf378a64d019c93ee2ac412830b2022-12-21T18:38:12ZengBMCEuropean Journal of Medical Research2047-783X2020-10-0125111210.1186/s40001-020-00453-yFibroblast-like cells change gene expression of bone remodelling markers in transwell culturesEliza S. Hartmann0Sabine Schluessel1Miriam I. Köhler2Felicitas Beck3Julia I. Redeker4Burkhard Summer5Veronika Schönitzer6Andreas Fottner7Susanne Mayer-Wagner8Department of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU MunichDepartment of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU MunichDepartment of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU MunichDepartment of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU MunichDepartment of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU MunichDepartment of Dermatology, Ludwig-Maximilians-UniversityDepartment of Surgery, Experimental Surgery, and Regenerative Medicine, Ludwig-Maximilians-UniversityDepartment of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU MunichDepartment of Orthopaedics, Physical Medicine and Rehabilitation, University Hospital, LMU MunichAbstract Introduction Periprosthetic fibroblast-like cells (PPFs) play an important role in aseptic loosening of arthroplasties. Various studies have examined PPF behavior in monolayer culture systems. However, the periprosthetic tissue is a three-dimensional (3D) mesh, which allows the cells to interact in a multidirectional way. The expression of bone remodeling markers of fibroblast-like cells in a multilayer environment changes significantly versus monolayer cultures without the addition of particles or cytokine stimulation. Gene expression of bone remodeling markers was therefore compared in fibroblast-like cells from different origins and dermal fibroblasts under transwell culture conditions versus monolayer cultures. Methods PPFs from periprosthetic tissues (n = 12), osteoarthritic (OA) synovial fibroblast-like cells (SFs) (n = 6), and dermal fibroblasts (DFs) were cultured in monolayer (density 5.5 × 103/cm2) or multilayer cultures (density 8.5 × 105/cm2) for 10 or 21 days. Cultures were examined via histology, TRAP staining, immunohistochemistry (anti-S100a4), and quantitative real-time PCR. Results Fibroblast-like cells (PPFs/SFs) and dermal fibroblasts significantly increased the expression of RANKL and significantly decreased the expression of ALP, COL1A1, and OPG in multilayer cultures. PPFs and SFs in multilayer cultures further showed a higher expression of cathepsin K, MMP-13, and TNF-α. In multilayer PPF cultures, the mRNA level of TRAP was also found to be significantly increased. Conclusion The multilayer cultures are able to induce significant expression changes in fibroblast-like cells depending on the nature of cellular origin without the addition of any further stimulus. This system might be a useful tool to get more in vivo like results regarding fibroblast-like cell cultures.http://link.springer.com/article/10.1186/s40001-020-00453-yImplant materialFibroblast-like cellsTranswell cultureBone remodelingAseptic loosening |
| spellingShingle | Eliza S. Hartmann Sabine Schluessel Miriam I. Köhler Felicitas Beck Julia I. Redeker Burkhard Summer Veronika Schönitzer Andreas Fottner Susanne Mayer-Wagner Fibroblast-like cells change gene expression of bone remodelling markers in transwell cultures European Journal of Medical Research Implant material Fibroblast-like cells Transwell culture Bone remodeling Aseptic loosening |
| title | Fibroblast-like cells change gene expression of bone remodelling markers in transwell cultures |
| title_full | Fibroblast-like cells change gene expression of bone remodelling markers in transwell cultures |
| title_fullStr | Fibroblast-like cells change gene expression of bone remodelling markers in transwell cultures |
| title_full_unstemmed | Fibroblast-like cells change gene expression of bone remodelling markers in transwell cultures |
| title_short | Fibroblast-like cells change gene expression of bone remodelling markers in transwell cultures |
| title_sort | fibroblast like cells change gene expression of bone remodelling markers in transwell cultures |
| topic | Implant material Fibroblast-like cells Transwell culture Bone remodeling Aseptic loosening |
| url | http://link.springer.com/article/10.1186/s40001-020-00453-y |
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